We propose to form a virtual Center for Protein Folding Machinery with 9 established investigators with complementary expertise in biology, chemistry, physics and computing from 5 academic institutions. Our goal is to define the basic chemical and physical principles used by molecular chaperones to promote the folding of key cellular proteins. This knowledge will form the basis of the applications for engineering protein machines that can assist the folding of any protein of interest and for developing strategies to alleviative or prevent protein misfolding associated with a number of human diseases. We choose the eukaryotic chaperonin TRiC (approximately 1 MDa) and the closely related chaperonin from M. maripaludis as our model systems because of their biomedical relevance and ease of purification and genetic modifications which will facilitate its physical characterizations. We plan to take an integrated approach to study these chaperonins with the most suitable physical and computational methods including electron cryomicroscopy, single-molecule spectroscopy and imaging, structure-constrained homology modeling and in-silico protein folding within the chaperonin environment. These various methodologies will need to be improved in order to address all aspects of this relatively large and structurally dynamic machinery. Consequently, our studies will also create new approaches applicable for study of biological machines in general. To integrate the various strengths of the individual research groups, we will develop flexible database and tele-communication schemes to communicate and exchange the diverse data produced in the Center. This planning grant will allow us to learn the merits of each technique and determine how they can best complement each other with the goal of developing a comprehensive research Center. We will conduct workshops, tele-conferences and various individual or group interactions. An external advisory panel will be chosen to guide us through the process of defining realistic goals, setting major milestones and implementing cross-institutional collaborations logistics.